Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation

Abstract

Using two-colour flow cytometry >200 antibodies submitted to the 8 th International Workshop of Human Leukocyte Differentiation Antigens (HLDA8) have been analyzed for their reactivity with resting and activated CD203c + basophils. Four antibodies either non-reactive or weakly reactive with resting basophils exhibited an increased reactivity with basophils activated by anti-IgE-mediated cross-linking of the high affinity IgE receptor (FcεRI). These include antibodies against CD164 (WS-80160, clone N6B6 and WS-80162, clone 67D2), as well as two reagents with previously unknown specificities that were identified as CD13 (WS-80274, clone A8) and CD107a (WS-80280, clone E63-880). The activation patterns followed either the "CD203c-like" or "CD63-like" activation profile. The CD203c profile is characterized by a rapid and significant upregulation (of CD13, CD164, and CD203c), reaching maximum levels after 5-15 min of stimulation. The phosphoinositide-3-kinase (PI3K)-specific inhibitor wortmannin inhibited the upregulation of these markers whereas 12-O-tetradecanoyl-phorbol-13-acetate (TPA) induced a rapid and FcεRI-independent upregulation within 1-2 min. In the CD63 profile, maximum upregulation (of CD63 and CD107a) was detected only after 20-40 min, and upregulation by TPA reached maximum levels after 60 min. In summary, our data identify CD13, CD107a, and CD164 as novel basophil-activation antigens. Based on time kinetics of upregulation, we hypothesize that molecules of the "CD203c group" and the "CD63 group" are linked to two different mechanisms of basophil activation.

Mechanism of phosphatidylserine inhibition of IgE/FcεRI-dependent anaphylactic human basophil degranulation via CD300a

“…Two different histamine-containing compartments, which show distinct early signaling requirements and kinetics, have been described, namely, the CD203c compartment and the CD63 compartment. 2,3 The appearance of CD63 is invariably associated with AND. 2 Mechanisms that regulate basophil activation likely relate to activation conditions (IgE/FcεRI-dependent and -independent), mode of degranulation (piecemeal degranulation vs AND), and changes in functional properties of inhibitory receptors.…”

CD300a is expressed on human basophils and seems to inhibit IgE/FcεRI‐dependent anaphylactic degranulation

“…The surface markers CD203c and CD63 were studied as readouts for basophil activation since they have been reported that up-regulation of these two molecules follows two distinct pathways (23).…”

A comparison between two different in vitro basophil activation tests for gluten- and cow’s milk protein sensitivity in irritable bowel syndrome (IBS)-like patients

“…This could be an important factor in the patients we studied as the presence of anti-gliadin IgG antibodies in 50% -60% of gluten-sensitive, IBS-like patients has been demonstrated [ 10 , 16 ] and we have demonstrated an evident overlap between patients " classified as gluten-sensitive " and those " classified as suffering from wheat hypersensitivity " [ 10 ]. Another factor which could negatively interfere with the accuracy of the new test is the reduced incubation time with the allergens, compared to the old test on separate leukocytes (15 min vs. 60 min) [ 17 ]. Finally, a direct comparison between the technique on whole blood and that on isolated leukocytes has been reported and the latter appeared to be more sensitive for allergen-induced basophil CD63 activation [ 18 ].…”